Tissue Eng Regen Med.
2017 Apr;14(2):179-185. 10.1007/s13770-017-0031-8.
Effects of clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) protein 9 system-Based Deletion of miR-451 in Mouse Embryonic Stem Cells on Their Self-Renewal and Hematopoietic Differentiation
- Affiliations
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- 1Department of Biomedical Science, Graduate School of Biomedical Science and Engineering, Hanyang University, 222 Wangsimni-ro, Seongdong-gu, Seoul 04763, South Korea. ks66kim@hanyang.ac.kr
- 2College of Medicine, Hanyang University, Seoul, South Korea.
- 3Department of Internal Medicine, School of Medicine, Kangwon National University, Kangwondaehakgil-1, Chuncheon, Gangwon-do 24341, South Korea. shhong@kangwon.ac.kr
- 4Department of Molecular and Cellular Biochemistry, School of Medicine, Kangwon National University, Chuncheon 24341, South Korea.
- 5Stem Cell Institute, Kangwon National University, Chuncheon 24341, South Korea.
- KMID: 2377348
- DOI: http://doi.org/10.1007/s13770-017-0031-8
Abstract
- Pluripotent stem cells (PSCs) are a useful source of cells for exploring the role of genes related with early developmental processes and specific diseases due to their ability to differentiate into all somatic cell types. Recently, the clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated (Cas) protein 9 system has proven to be a robust tool for targeted genetic modification. Here, we generated miR-451-deficient PSCs using the CRISPR/Cas9 system with PCR-based homologous recombination donor and investigated the impact of its deletion on self-renewal and hematopoietic development. CRISPR/Cas9-mediated miR-451 knockout did not alter the gene expressions of pluripotency, cellular morphology, and cell cycle, but led to impaired erythrocyte development. These findings propose that a combination of PSCs and CRISPR/Cas9 system could be useful to promote biomedical applications of PSCs by elucidating the function and manipulating of specific miRNAs during lineage specification and commitment.
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